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  • Induction of murine TNBS colitis is strictly controlled by a modified method using continuous inhalation anesthesia with sevoflurane.

Induction of murine TNBS colitis is strictly controlled by a modified method using continuous inhalation anesthesia with sevoflurane.

Digestive diseases and sciences (2014-01-24)
Tomohiro Terai, Satoshi Osawa, Shinya Tani, Shinji Oishi, Yoshifumi Arai, Takanori Yamada, Mitsushige Sugimoto, Takahisa Furuta, Shigeru Kanaoka, Hiroaki Miyajima, Ken Sugimoto
ABSTRACT

Trinitrobenzenesulfonic acid (TNBS)-induced colitis is one of the most widely used experimental colitis models. However, there is no standard procedure for inducing colitis by TNBS because it is difficult to achieve a uniform distribution of colitis. We have developed a modified method of murine TNBS-induced colitis that involves inhalation anesthesia with sevoflurane combined with both single and repeated TNBS administrations. To compare the usefulness of our newly developed method for inducing murine TNBS-induced colitis with that of conventional intraperitoneal anesthesia. TNBS in ethanol was administered to C57BL/6J mice held in an inverted vertical position either under continuous inhalation anesthesia with sevoflurane, in accordance with our newly developed method, or by intraperitoneal injection with 2.5 % avertin, in accordance with the conventional procedure. Body weight change, cytokine profile, and histological findings were examined during the course of colitis. The dispersion of anesthesia time, TNBS retention time, and nadir weight during the course of colitis was decreased using the newly developed method compared with the conventional procedure. Optimization of the modified TNBS-induced colitis, as evidenced by the predominant expression of Th1 and Th17 cytokines on day 7, was attained by the injection of 2.25 mg TNBS in 55 % ethanol. Regulation of the TNBS retention time using inhalation anesthesia with sevoflurane allowed strict control of the disease severity of TNBS-induced colitis. Using the modified method we were also able to develop a chronic TNBS-induced colitis model by repeated TNBS administration without excessive mortality of the mice. Our modified method for murine TNBS-induced colitis using continuous inhalation anesthesia with sevoflurane provides a better experimental colitis model following both single and repeated TNBS administrations.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Picrylsulfonic acid solution, 5 % (w/v) in H2O, BioReagent, suitable for determination of primary amines
Sigma-Aldrich
2-Methyl-2-butanol, ReagentPlus®, 99%
Sigma-Aldrich
Picrylsulfonic acid solution, 1 M in H2O
Sigma-Aldrich
2-Methyl-2-butanol, ReagentPlus®, ≥99%
Sigma-Aldrich
2-Methyl-2-butanol, anhydrous, ≥99%
Supelco
2-Methyl-2-butanol, analytical standard
Sevoflurane, European Pharmacopoeia (EP) Reference Standard